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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K1199044 SequenceAndFeatures</partinfo>
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==Improvement from UESTC-China==
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As a mutant with high activity toward 1,3-DCP, HheC-W249P has little change in the activity of 2,3-DCP. This year we UESTC-China used this part as the substrate to successfully obtain two mutants BBa_K2286006 (P84A) and BBa_K2286008 (P84-W249P) with improved 2,3-DCP activity by molecular simulation and single-site saturation mutagenesis.
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===Activity toward 2，3-DCP===
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Halogen ion release occurs when the haloalcohol dehalogenase catalyzes the the o-halanol to the epoxide. The interaction of the halide with Hg2+ allows Hg2+ to be separated from SCN-, which forms a colored complex with Fe3 +. This complex can be determined by absorption at 460 nm. Therefore, we determined these two mutants activity by the halide ion method.
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              <p>Figure 1. The activity of mutants on 2,3-DCP</p>
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              <p>Data were measured in 50mM Tris-H2SO4 at pH 8.5 and 37℃</p>
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Revision as of 07:51, 30 October 2017

HheCW249P 2,3-DCP(2,3-dichloropropanol)->Glycerol

HheC displayed high regioselectivity and moderate to high enantioselectivity that can be applied for the kinetic resolution of chiral spiroepoxides.And HheC-W249P is mutant of HheC ,displayed higher activity than the wide type .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 697
  • 1000
    COMPATIBLE WITH RFC[1000]

Improvement from UESTC-China

As a mutant with high activity toward 1,3-DCP, HheC-W249P has little change in the activity of 2,3-DCP. This year we UESTC-China used this part as the substrate to successfully obtain two mutants BBa_K2286006 (P84A) and BBa_K2286008 (P84-W249P) with improved 2,3-DCP activity by molecular simulation and single-site saturation mutagenesis.

Activity toward 2，3-DCP

Halogen ion release occurs when the haloalcohol dehalogenase catalyzes the the o-halanol to the epoxide. The interaction of the halide with Hg2+ allows Hg2+ to be separated from SCN-, which forms a colored complex with Fe3 +. This complex can be determined by absorption at 460 nm. Therefore, we determined these two mutants activity by the halide ion method.

Figure 1. The activity of mutants on 2,3-DCP

Data were measured in 50mM Tris-H2SO4 at pH 8.5 and 37℃